Single-molecule fluorescence spectroscopy offers real-time, nanometer-resolution information. Over the past two decades, this emerging single-molecule technique has been rapidly adopted to investigate the structural dynamics and biological functions of proteins. Despite this remarkable achievement, single-molecule fluorescence techniques must be extended to macromolecular protein complexes that are physiologically more relevant for functional studies. In this review, we present recent major breakthroughs for investigating protein complexes within cell extracts using single-molecule fluorescence. We outline the challenges, future prospects and potential applications of these new single-molecule fluorescence techniques in biological and clinical research.
Bibliographical noteFunding Information:
C.J. was supported by Starting Grants (ERC-StG-2012-309509) through the European Research Council. M.F. was supported by Fondation pour la Recherche Medicale. V.N. was supported by the Research Center Program (EM1202) of IBS (Institute for Basic Science), and the National Honor Scientist Program (20100020415) through the National Research Foundation of Korea (NRF). We appreciate helpful discussions with Jeff Gelles and Antoine van Oijen. We also appreciate critical comments from Elio Abbondanzieri, Timothy Blosser, Stanley Chandradoss, Jetty van Ginkel, Anna Haagsma, Inha Heo, Sungchul Hohng, Kaushik Ragunathan, and Kyu-Hyeon Yeom. We apologize for being unable to cite other research papers and reviews owing to space limitations.
- Cell lysate
- Macromolecular complex
- Single-molecule fluorescence
- Single-molecule immunoprecipitation
- Single-molecule pull-down